myceliophthora thermophila wild type strain atcc  (ATCC)

Journal: Biology

Article Title: Development of Genetic Tools in Glucoamylase-Hyperproducing Industrial Aspergillus niger Strains

doi: 10.3390/biology11101396

Figure Lengend Snippet: Transformation of A. niger N1 and O1 was carried out by protoplast-mediated transformation (PMT) using the vector of pAN52-P ahr - gfp -T ahr ( A ) and Agrobacterium tumefaciens -mediated transformation (AMT) using the vector of pPK2- hph - gfp ( B ). pAN52-P ahr - gfp -T ahr was transformed into protoplasts of A. niger N1 and O1 by PMT. The pPK2- hph-gfp was transformed into conidia of A. niger N1 and young mycelia of A. niger O1 by AMT. P ahr is the promoter of the alkyl hydroperoxide reductase from M. thermophila ; P tef1 is the promoter of the translation elongation factor gene tef1 from A. niger ; P trpC is the promoter of the tryptophan synthetase gene from A. nidulans ; GFP, enhanced green fluorescence protein; Neo , neomycin-resistance gene; Hph , hygromycin resistance gene; RB and LB, right and left border of T-DNA, respectively.

Article Snippet: In previous studies of PMT, 10 8 conidia from Aspergillus giganteus strain IfGB 15/0903 and Myceliophthora thermophila wild-type strain ATCC 42,464 were incubated in liquid culture to generate a sufficient yield of protoplasts, with 55 and 25 transformants being obtained in A. giganteus and M. thermophila , respectively [ , ].

Techniques: Transformation Assay, Plasmid Preparation, Fluorescence

Journal: Biotechnology for Biofuels

Article Title: Development of a genome-editing CRISPR/Cas9 system in thermophilic fungal Myceliophthora species and its application to hyper-cellulase production strain engineering

doi: 10.1186/s13068-016-0693-9

Figure Lengend Snippet: Phenotypic analysis of constitutive Cas9-expressing strains. a Fluorescence microscopic assessment of Cas9-gfp localization in M. thermophila . The nuclei were stained with 4′,6-diamidino-2-phenylindole. Each scale bar represents 10 µm. b Colony growth and sporulation of Cas9OE and wild-type (WT) strains on minimal medium plates after 4 days of culture

Article Snippet: The genome of the M. thermophila wild-type (WT) strain ATCC 42464 has been completely sequenced and annotated [ ], thereby allowing systematic examination and identification of lignocellulolytic enzymes cultured in defined plant-derived biomass through transcriptome and exoproteome approaches [ , ].

Techniques: Expressing, Fluorescence, Staining

Journal: Biotechnology for Biofuels

Article Title: Development of a genome-editing CRISPR/Cas9 system in thermophilic fungal Myceliophthora species and its application to hyper-cellulase production strain engineering

doi: 10.1186/s13068-016-0693-9

Figure Lengend Snippet: Introduction of directed mutations into the amdS gene in M. thermophila by sgRNA-guided Cas9. a Schematic illustration of amdS mutagenesis by the CRISPR/Cas9 system. The M1 strain carrying the amdS gene was co-transformed with U6p-amdS-sgRNA and Cas9 cassettes, and then positive transformants were selected on medium plates containing 2 mg/mL FAA. b Sequence alignment of the amdS target locus from FAA-resistant transformants. The wild-type target sequence of amdS is framed in blue . Red letters depict the protospacer adjacent motif (PAM)

Article Snippet: The genome of the M. thermophila wild-type (WT) strain ATCC 42464 has been completely sequenced and annotated [ ], thereby allowing systematic examination and identification of lignocellulolytic enzymes cultured in defined plant-derived biomass through transcriptome and exoproteome approaches [ , ].

Techniques: Mutagenesis, CRISPR, Transformation Assay, Sequencing

Journal: Biotechnology for Biofuels

Article Title: Development of a genome-editing CRISPR/Cas9 system in thermophilic fungal Myceliophthora species and its application to hyper-cellulase production strain engineering

doi: 10.1186/s13068-016-0693-9

Figure Lengend Snippet: Phenotypic analysis of Δcre-1 and wild-type strains. a Colonies of Δ cre - 1 (Δ Mtcre - 1 and Δ Mhcre - 1 ) and wild-type strains (MtWT and MhWT) of M. thermophila and M. heterothallica on minimal medium plates after 2 days. b Sodium dodecylsulfate-polyacrylamide gel electrophoresis of secreted protein of Δ cre - 1 and wild-type strains after 4 days culture in 2% Avicel medium. c – f Assays for protein concentration and xylanase, endoglucanase, and exoglucanase activities of Δ cre - 1 and wild-type strains in 2% Avicel inducing medium after 3 and 4 days culture. Bars marked by asterisks in each group differ significantly from unmarked ones (Tukey’s HSD, p < 0.001). Error bars represent SD from three replicates

Article Snippet: The genome of the M. thermophila wild-type (WT) strain ATCC 42464 has been completely sequenced and annotated [ ], thereby allowing systematic examination and identification of lignocellulolytic enzymes cultured in defined plant-derived biomass through transcriptome and exoproteome approaches [ , ].

Techniques: Polyacrylamide Gel Electrophoresis, Protein Concentration

Journal: Biotechnology for Biofuels

Article Title: Development of a genome-editing CRISPR/Cas9 system in thermophilic fungal Myceliophthora species and its application to hyper-cellulase production strain engineering

doi: 10.1186/s13068-016-0693-9

Figure Lengend Snippet: CRISPR/Cas9-mediated homologous recombination (HR) efficiency of simultaneous disruption of one to four gene loci

Article Snippet: The genome of the M. thermophila wild-type (WT) strain ATCC 42464 has been completely sequenced and annotated [ ], thereby allowing systematic examination and identification of lignocellulolytic enzymes cultured in defined plant-derived biomass through transcriptome and exoproteome approaches [ , ].

Techniques: CRISPR, Homologous Recombination